NESTIN (HU); MSX - 100UG

Stock Code: 3572043
Manufacturer Part No: MAB5326
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Analysis Note


ControlPositive Control: WI-38 (ATCC# CCL75) or U251 cell lines


Application


Research CategoryStem Cell Research


Research Sub CategoryNeural Stem Cells


This Anti-Nestin Antibody, clone 10C2 is validated for use in IC, IH, IH(P), WB for the detection of Nestin.


Immunoblotting: 1:1,000-1:2,000 on denatured protein. Western blot: 1:1000-1:5000. Recognizes a 220-240kDA band on denaturing SDS gels. There is also a 60-64kDa band present in most lysates, the identity of this smaller fragment is unknown but it is not seen in lysates not containing human nestin protein.

WI-38 (ATCC# CCL75) can be used as a positive control. U251 cell line is also positive.

Immunocytochemistry: 1:200 on cells fixed in 4% paraformaldehyde and permeabilized with 0.02% Triton

Immunohistochemistry: paraformaldehyde is the preferred fixative: 1:200 on formalin fixed, paraffin embedded tissue sections. Use triton 0.02% in block only; dilution buffer 0.1% BSA; incubations 1-2 hours RT. Human fetal brain is suggested as a positive control. (CA Messam et al, Develop. Brain Res, 134:87-92, 2002; Gu, et al., Brain Res. 943:174-180, 2002). Filamentous staining in the cytoplasm of cells

Optimal working dilutions must be determined by end user.


Disclaimer


Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.


General description


Nestin, a large intermediate filament protein (class Type VI) expressed during development and in myotendinous and neuromuscular junctions. Nestin expression is restricted, typically disappearing by E18. Nestin is thought to be a reasonable neuronal marker; however, recent studies have found nestin expression in other cell types such as endothelial cells {Folia Biologica, 44(5):155, 1998}. Nestin identifies the most primitive neuroepithelium but also identifies many other embryonic tissues, so it is not specific for CNS. Nestin expression is seen in almost all GBMs (Glioblastoma mulitformes) and many melanomas (both primary and metastatic) but not in any metastatic carcinoma.


Immunogen


Epitope: human


Fusion protein.


Legal Information


CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany


Linkage


Replaces: AB5922


Other Notes


Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.


Physical form


Protein A purified


Format: Purified


Protein A Purified mouse immunoglobulin in 20 mM sodium phosphate, 250 mM NaCl, pH. 7.6, with 0.1% sodium azide as a preservative.


Specificity


Reacts with human Nestin, with weak reactivity to monkey nestin. Not reactive with rodent nestin.


Storage and Stability


Antibodies may be stored at 4°C or -20°C. Avoid repeated freeze/thaw cycles.

ManufacturerSIGMA-ALDRICH

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